single-cell RNA sequencing on Drosophila melanogaster brain in an ALS/FTD model
single-cell RNA sequencing on Drosophila melanogaster brain in an ALS/FTD model
Fly dissection on ice for less than 1.5 hr. Collect the brains in PBS. Centrifuge the brains at 2000 rpm at 4 degree C for 5 minutes. Substitute the supernatant PBS with 50 uL dispase (3mg/ml) and 75 ul collagenase (100mg/ml). Incubate the brains with the enzymes at 25 degree C in a 500 rpm shaker for 5 minutes. Pipetting the enzymes solution containing the brains for another 10 minutes to fully dissociate brains. Centrifuge the dissociated cells at 4 degree C 2000 rpm for 5 minutes and wash the cells with 200 ul 0.04% BSA in PBS. Pre-wet the 10 um cell strainer. Pass the cell suspension through the strainer and collect the filtered cell suspension, and the sample is ready for viability check. 10X chromium 3' 10X chromium 3'